UA146658U - METHOD FOR DETERMINATION OF CLEANED TUBERCULINE ACTIVITY FOR MAMMALS IN BOVINE ANIMALS - Google Patents

Abstract

Method for determining purified tuberculin activity (PPD) for mammals in cattle vaccinated with live culture of mycobacteria of non-pathogenic BCG strain. BCG mycobacterial suspensions are administered twice, which makes it possible to use sensitized animals to control tuberculin activity for 3 months.

Description

A useful model relates to immunology, in particular to determining the activity of purified tuberculin (PPD) for mammals and can be used to monitor the activity of commercial batches of tuberculin produced under biological factory conditions. Purified tuberculin (PPD) for mammals is a derivative of pure protein (Rigiiea Rgoiviiip Oeegimaiime).

There is a method of controlling the activity of tuberculin (PPD) for mammals on cattle suffering from tuberculosis (Technical conditions of the State Standard of the USSR GOST 16739-88 Moscow, p. 21). However, this method has disadvantages. In particular, it is performed on cattle suffering from tuberculosis, which require separate premises and special conditions that meet the requirements of epizootic and epidemic safety.

The closest analogue of the useful model is the Method for determining the activity of purified tuberculin (PPD) for mammals (Patent Ukraine Mo 36006, class AbIK 39/04, dated 10.12.1999). The disadvantage of this method is that animals sensitized by suspension of the BCG vaccine strain culture are used once when determining the biological activity of only one production series of tuberculin.

The basis of a useful model is the task of developing a method for determining the activity of purified tuberculin (PPD) for mammals on cattle inoculated with a live culture of mycobacteria of a non-pathogenic strain of BCG by means of a two-time administration of a suspension of BCG mycobacteria, which makes it possible to use sensitized animals to control the activity of tuberculin for 3- x months

A comparative analysis with the closest analogue allows us to conclude that the introduction of BCG suspension twice, with an interval of 30 days, makes it possible to use these animals to determine the activity of tuberculin within 3 months and to determine the biological activity of 3 commercial series of tuberculin for mammals, which meets the criterion "novelty".

The method is performed as follows.

In a tuberculosis-free household, 24 animals are selected from the fattening group. healthy, tuberculin-resistant cattle aged 6 months or more.

The selected animals are divided into two groups of 12 heads each. Animals of the first group are injected with a suspension of a live culture of mycobacteria of the BCG vaccine strain once subcutaneously, and

Animals of the second group are injected twice (subcutaneously and intramuscularly). Determination of the activity of commercial series of tuberculin (PPD) for mammals, produced at a biological enterprise, is carried out in comparison with a control series of tuberculin.

Example Mo 1 (table 1). 30 days after the introduction of a live culture suspension of mycobacteria of the BCG vaccine strain, 24 animals were intradermally injected with purified mammalian tuberculin (PPD) in a standard solution of a commercial (production) series from the left side within the middle third of the neck with a needle-free injector, and from the right side - purified tuberculin (PPD) for mammals of the control series in a dose of 0.1 cmU in previously cut and treated with 70" ethyl alcohol.

Reactions in animals were recorded 72 hours after administration of tuberculins and assessed by the degree of thickening of the skin fold at the site of tuberculin injection in (mm) using a cuitimeter only in animals that reacted to both allergens with thickening of the skin fold by 3 mm or more. The obtained results are included in the report (Appendix 1). When determining the biological activity of production batches of tuberculin, there should be at least 10 heads that react positively to both allergens. At the same time, only animals that responded positively to the test and control series of tuberculin were taken into account for the account of reactions to tuberculin. Animals that reacted with the same intensity of reactions to both allergens were not taken into account.

The difference in intensity of reactions to experimental and control series of purified tuberculin (PPD) for mammals was determined using the table (Appendix 2).

An experimental series of tuberculin produced at a bioenterprise is considered active if there is no difference in the intensity of tuberculin reactions or it is statistically unreliable.

Table 1 (example 1) shows that after 30 days among the animals vaccinated with a suspension of live culture of BCG, which reacted to tuberculin of the experimental series, in comparison with the control series of tuberculin, they reacted with a greater intensity of reactions (tk) 4 gol., with a lower intensity (-) 6 goals. and with the same intensity (-) 2 goals... 60 days after the introduction of tuberculins, the intensity of reactions in animals to allergens decreased. At the same time, they reacted with a greater intensity of reactions (w) 4 heads, with a lower intensity (-) With heads. and with the same intensity (-) 1 goal... Thickening of the skin fold by 2 mm was noted in 2 animals and allergic reactions occurred in 2 animals. After 60 days, 2 patients reacted with a greater intensity of reactions (Ж), with a lower intensity (-) in 2 patients, with the same intensity (-) in 1 patient, a thickening of the skin fold by 2 mm was observed in 2 patients. and negative reactions in 5 goals...

Example Mo 2 (table 2). Also, as in example Mo 1, only the determination of tuberculin activity is carried out on cattle vaccinated with BCG vaccine strain culture twice.

From the materials in Table 2, it can be seen that 30 days after the subcutaneous injection of suspended live cultures of mycobacteria pcs. BCG intradermal reactions to tuberculin of experimental and control series were noted in 12 animals, but the intensity of reactions was different. Thus, in comparison with the control series, tuberculin of the experimental series reacted with a greater intensity of reactions (9 4 subjects, with a lower intensity (-) 6 subjects and with the same intensity (:-) 2 subjects... After 60 days, they reacted with a greater intensity of reactions (w) 4 subjects, with a lower intensity (-) 7 subjects and with the same intensity (-) 1 subject... After 90 days, they reacted with a greater intensity of reactions (w)

With goals, with less intensity (-) 7 goals. and with the same intensity (:-) 2 goals...

On the basis of the conducted studies of the commercial (research) series of tuberculin, in comparison with the control series of tuberculin, in experiments on healthy cattle vaccinated twice with a live culture of BCG mycobacteria, it was determined that the research (commercial) series of tuberculin is active and suitable for allergic diagnosis of tuberculosis agricultural animals.

The results of the conducted research indicate that the developed method is specific and safe for the health of animals and humans and allows determining the biological activity of commercial (production) series of purified tuberculin (PPD) for mammals on the same experimental animals within 90 days. This method is 4-5 times more economical, as well as simple in practice and safe in epizootic and epidemic terms.

Table 1

A method for determining the activity of purified tuberculin (PPD) for mammals

Mo one-time pcs. BCG due to (days) or the same (-) reaction to tva- . . . experimental and control series | 60 | 90 | 30 | 60 | 90 / z0 | 60 | 90 11 0 19151 | 70 5 | -1- I- 2 | 12 | lol | with | lo | 9 | 4 | xx | - | - from 7 | 5141-1814 1 - | - | I | negative 4 | 6 | 2 1 - 161 2 | - |- |negative | negative 5 8 | 5 1 219 | 5 1 2 | - | - |negative 61 9 | 6 | from 071 4 her her 7 5 1-1 - 17-11 - | - negative | negative 8 6 | - 1-15 1 - 1 - | I |negative |negative 91 12 | 8 | 4 | n 6 | with | 5 | x- | with lo |! lo | 5 | 2 | lo | 5 | 2 | |negative |negative m ml 115-172 | 6 | - | - | - |negative

Table 2

A method for determining the activity of purified tuberculin (PPD) for mammals

Mo twice sh... BCG through (days) or the same (-) reaction to tva- . . . experimental and control series | in | 90 | 30 | 60 | 90 | from | in | 980 2 | 15 | 718 | 12 | 13 | 17 | 714 | 5 | - | - 3 | 13 | 15 | 13 | 14 | 18 | 74 | - | - | - 4 | 710 | with | 13 | 12 | 15 | 74 | - | - | - 6 13 | 15 | 13| 14 | 17 | 15 | - HER - |/ - 7 | 10 | 17 | 1519 | t12| 70 | 5 | In | h- 8 | 10 | 714 | 12 | 12 | 16 | 712 | | 1 - 9 8 | 712 |t10| 9 | 15 "h |- her - - 710! 9 | 131919 | 131 9 |- |- |- 117 710 | 712 | 9 | lo | 15 | l70 |I- | - | - 12| 15 | h |15| 18 | 24 | 718 | - | - | -

Appendix 1

A method for determining the activity of purified tuberculin (PPD) for mammals

The presence of larger (sq. m), about animals. . and M and

Experimental series Control series of the reaction to the experimental and control series of tuberculin pi p PT I PO: POLON NO nI"?"n;ESIISHІІІ IOV. TL OTTO TE S NO chinchillas i LY NI I shi i i - o PO: POLYA PO 77111118 p i G.YA PO: POLON NO shi pn I V: TO PO 27071779 111111711171171111111111712 1 negative reaction.id//:/ py ka PO: TO PO: ON NO p R PL: TI PO SPON NO

Appendix 2

The method of determining the activity of purified tuberculin (PPD) for mammals c, . : : Maximum number of animals with greater (x) or

The number of animals with different intensity and . also: y and - less (-) intensity of reactions to the experimental reactions to the experimental and control series. b. May and tuberculin series of tuberculin, according to which withdrawal from control is probable (95-97.5 Uo nn"n:ШШЙ nn

Claims (1)

USEFUL MODEL FORMULA A method for determining the activity of purified tuberculin (PPD) for mammals in cattle inoculated with a live culture of mycobacteria of a non-pathogenic strain of BCG, which is distinguished by the fact that a suspension of mycobacteria BCG is administered twice, which makes it possible to use sensitized animals to monitor the activity of tuberculin for 3 x months